WO2017061787A1 - Composition for relieving hangovers containing extracellular polysaccharide produced from ceriporia lacerata as active ingredient - Google Patents

Abstract

The present invention relates to extracellular polysaccharide produced from Ceriporia lacerata, a mycelium culture medium of Ceriporia lacerata containing the extracellular polysaccharide, and a composition for relieving hangovers containing dry powder or an extract of the mycelium culture medium as an active ingredient. The composition according to the present invention reduces blood ethanol or acetaldehyde level and is thus excellent in relieving hangovers.

Description

Hangover relief composition containing extracellular polysaccharide produced by Seriphoria laccerata as an active ingredient

The present invention is an active ingredient produced by Ceriporia lacerata ( Ceriporia lacerata ), the mycelium culture medium of the Ceriporia laccerata containing the active ingredient, the dry powder of the mycelium culture solution or the extract of the mycelium culture medium active ingredient It relates to a composition for hangover relief to contain.

In general, "hangover" refers to a phenomenon in which the head is heavy, ill or sore due to drinking, which is caused by the poisoning effect of ethanol or acetaldehyde accumulated in hepatocytes. . If these substances accumulate in liver cells for a long time and cause poisoning, fatigue, abdominal bloating, and vomiting may occur.

In normal ethylalcohol metabolism, the ethyl alcohol introduced into the body is absorbed by the gastrointestinal or small intestine and enters the blood vessels and is transferred to the liver. Meanwhile, hepatocytes have alcohol dehydrogenase (ADH) to oxidize alcohol to acetaldehyde, and the acetaldehyde is decomposed to acetic acid by acetaldehyde dehydrogenase (ALDH) in hepatocytes. The decomposed product is transferred to muscle or adipose tissue of the whole body and finally decomposed into carbon dioxide gas and water.

Acetaldehyde dehydrogenases include type II which initiates oxidation even at low concentrations of acetaldehyde and type I which initiates oxidation only at high concentrations of acetaldehyde. Asians are generally deficient in type II acetaldehyde dehydrogenase, which slows the oxidation of acetaldehyde, and thus causes various hangovers due to normal metabolism caused by unoxidized acetaldehyde and / or ethyl alcohol. .

On the other hand, ethanol is the main ingredient of alcohol, ingested ethanol is absorbed through the small intestine and digestive tract, and blood alcohol concentration reaches its highest between 20 and 120 minutes after ingestion. This absorbed ethanol is metabolized in all organs, including the liver, about 10% of which is excreted in breathing, or in urine and sweat, and most of it is broken down in the liver.

Ethanol degradation in the liver is the main metabolism of conversion to acetaldehyde through oxidation. This is done by three reactive enzyme systems: ADH, microsomal ethanol-oxidizing system and catalase (K. Ebihara et al. , Agri . Biol . Chem . , 52, 1311, 1988). . As described above, various toxicological studies of ethanol have been conducted as well as toxicological studies. It has been reported that toxicities of ethanol not only affect neurological aspects but also genetics (J. Caballeria, et al. , Life Sci. , 41, 1021-1727, 1986).

In recent years, research and experiments have been conducted on a number of substances that can reduce the toxicity of ethanol or inhibit the expression of toxic substances. As a result, various health supplements containing ingredients extracted from natural or herbal ingredients have been developed. have. As part of this research, Korean Patent No. 10-0968109 discloses a hangover relief composition comprising a natural extract and a calcium compound and a method of manufacturing the same.

Ceriporia laccerata is a type of white fungus and is known to perform co-metabolism called lignin decomposition in order to use carbon sources such as cellulose, hemicellulose, other polysaccharides, and glycerol in an ecosystem.

Regarding the medical treatment using Ceriporia laccerata, only the use of diabetic treatment of culture extract of Ceriporia laccerata in Korean Patent No. 10-1031605 is known so far, and Ceriporia laccerata is known. Hangover use has not been reported yet.

Accordingly, the present inventors have completed the present invention by confirming that the mycelial culture solution, the dry powder or extract thereof, of the active ingredient isolated from Seriphoria laccerata, the same, and the Seriphoria laccerata, exhibit a hangover effect.

It is an object of the present invention to provide a pharmaceutical composition and health functional food for preventing or eliminating hangovers containing the active ingredient produced by Seriphoria laccerata.

It is another object of the present invention to provide a method for preventing or eliminating hangover using the active ingredient produced by Ceriporia laccerata.

Another object of the present invention is to provide the use of the active ingredient produced by Ceriporia laccerata for use in the manufacture of a pharmaceutical composition for preventing or relieving hangovers.

In order to achieve the above object, the present invention is an extracellular polysaccharide produced by Seriphoria laccerata, mycelium culture medium of the Seriphoria laccerata comprising the extracellular polysaccharide, dry powder of the mycelium culture medium or the mycelium culture medium It provides a pharmaceutical composition for preventing or resolving hangover, containing the extract of as an active ingredient.

In addition, the present invention is an extracellular polysaccharide produced by Seriphoria laccerata, mycelium culture medium of the Seriporia laccerata containing the extracellular polysaccharide, the dry powder of the mycelium culture medium or the extract of the mycelium culture medium active ingredient Provided as a health functional food for preventing or improving hangovers.

In addition, the present invention requires an extracellular polysaccharide produced by Seriphoria laccerata, mycelium culture medium of the Seriphoria laccerata containing the extracellular polysaccharide, a dry powder of the mycelium culture medium or an extract of the mycelium culture medium. It provides a hangover prevention or remedy comprising administering to a subject.

Furthermore, the present invention is an extracellular polysaccharide produced by Ceriporia laccerata for use in the manufacture of a pharmaceutical composition for preventing or eliminating hangovers, a mycelium culture solution of a Ceriporia laccerata comprising the extracellular polysaccharide, It provides a dry powder of the mycelium culture medium or the use of the extract of the mycelium culture medium.

The present invention contains an extracellular polysaccharide produced by Seriphoria laccerata, a mycelium culture medium of the Seriphoria laccerata containing the extracellular polysaccharide, a dry powder of the mycelium culture medium, or an extract of the mycelium culture medium as an active ingredient. The pharmaceutical composition and health functional food for preventing or eliminating hangovers exhibit an excellent hangover effect by reducing ethanol and / or acetaldehyde in the blood.

Figure 1 shows the results of measuring the blood alcohol concentration of the mice after treatment of the culture medium of the seriporia laccerata mycelium administered to the ethanol.

Figure 2 is a result of measuring the hangover resolution of the seriporia lacserata mycelium culture solution through a simple clinical experiment.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The present invention contains an extracellular polysaccharide produced by Seriphoria laccerata, a mycelium culture medium of the Seriphoria laccerata containing the extracellular polysaccharide, a dry powder of the mycelium culture medium or an extract of the mycelium culture medium as an active ingredient. To provide a pharmaceutical composition for preventing or alleviating hangovers.

As used herein, the term "extracellular polysaccharide (EPS)" is a part of the cell wall of a microorganism such as a fungus, and the polysaccharide is secreted to the outside of the cell to form a capillary around it or secreted into the periphery or medium as a slime. Means. The extracellular polysaccharide is secreted by the microorganism to protect itself from the external environment such as antibodies, toxic substances, protozoa and bacteriophage.

The extracellular polysaccharide is 40 to 60% by weight of sugar and 30 to 40% by weight of protein, 40 to 50% by weight of sugar and 32 to 38% by weight of protein, 43 to 47% by weight of sugar and 33 to 36% by weight Protein, and more specifically about 45% by weight of sugar and about 34% by weight of protein.

The sugar may contain mannose, galactose and glucose.

The extracellular polysaccharide may have a molecular weight of 100 ~ 150 kDa, 110 ~ 140 kDa or 115 ~ 125 kDa, more specifically may have a molecular weight of about 120 kDa.

In one embodiment of the present invention, the extracellular polysaccharide, (a) liquid culture of the mycelium of Seriporia laccerata to prepare a mycelium culture medium of Seriporia laccerata; (b) drying the powdered mycelium culture medium of the seriporia laccerata; And (c) extracting the dry powder of the mycelium culture medium of Seriphoria laccerata with a solvent, filtering the same, and concentrating under reduced pressure.

The medium for liquid culture of the mycelia of the seriporia laccerata in step (a) is sugar, glucose, starch, hydrate, soybean meal, soy flour, magnesium sulfate (MgSO ₄ ), potassium phosphate (KH ₂ PO). ₄ ), potassium diphosphate (K ₂ HPO ₄ ) and water, and the hydrogen ion concentration may be pH 4.5 ~ 6.0.

Specifically, the medium, 0.2 to 3% by weight of sugar, 0.2 to 3% by weight of glucose, 0.2 to 4% by weight, 0.1 to 0.5% by weight of water, 0.1 to 0.5% by weight of wheat flour, 0.2 to 3 weight of soy flour %, 0.01% to 0.1% by weight of magnesium sulfate (MgSO ₄ ), 0.01% to 0.25% by weight of potassium monophosphate (KH ₂ PO ₄ ), 0.01% to 0.25% by weight of potassium diphosphate (K ₂ HPO ₄ ) and the balance of water Can be.

Liquid culture in the step (a) may be carried out by maintaining the concentration of carbon dioxide under 1,000 to 2,000 ppm under a blue LED light source.

The liquid culture, for example, at 20 ~ 28 ℃ hydrogen ion concentration (pH) of 4.5 to 6.0, the light source to maintain a blue LED, 0.1 to 0.8 LUX illuminance, 0.5 to 2.0 kgf / ㎠ air injection The concentration of carbon dioxide may be performed for 8 to 13 days while maintaining the concentration at 1,000 to 2,000 ppm. Specifically, it may be performed for 5 to 15 days at the conditions of 20 ~ 25 ℃, pH 4.5 ~ 6.0, 0.5 ~ 2.0 kgf / ㎠, and 1,000 ~ 2,000 ppm. When the liquid culture under the conditions described above, the content of the extracellular polysaccharide produced is high.

The parent strain in the step (a) is about one excellent strain stored at 1 ~ 5 ℃ in PDA (Potato dextrose agar) medium state, using a PDB (Potato dextrose broth) medium in an Erlenmeyer flask, about 25 in shake incubator It can be used after incubating for 7-9 days while maintaining the temperature of ℃. In addition, the culture solution or mycelium obtained after culturing the parent strain can be used as the inoculum. Specifically, the amount of mycelium to be added to the inoculum may be about 0.5% (w / v) based on the amount of the solution to be cultured. The amount of extracellular polysaccharides does not increase as the amount of mycelia (% / 100 ml, w / v) increases, so the medium composition has the highest content of extracellular polysaccharides, not the best nutritional ratio and environmental conditions for the growth of mycelia. It is preferable to apply the selective culture conditions to form.

The culture solution can be separated and purified into a mycelium and an aqueous solution. For the separation and purification, the solution from which the mycelium is removed by centrifugation may be repeatedly purified using a multi-sheet filter press and a vibration centrifugal separator (PALLSEP), and then irradiated with ultraviolet (UV) light for 1 minute. In addition, the culture solution may be stored after sealing to remove oxygen, which may bring a change in the content of the active ingredient due to the growth of the mycelia when the mycelia exist in the culture solution.

In step (b), the mycelium culture solution prepared in step (a) may be dried and powdered. The drying may be performed for 48 to 96 hours at a temperature of 40 ℃ or less, more specifically 30 ℃ or less to prevent the disappearance of the active material. In addition, the drying is preferable to use a vacuum freeze dryer rather than a vacuum dryer to set the evaporation temperature relatively high because the change in the effective substance content is minimized.

In the step (c), after extracting the dry powder of the mycelium culture medium obtained in step (b) with a solvent, the extracellular polysaccharide which is an active ingredient of the composition according to the present invention is separated.

Specifically, 100 ml of distilled water was added to the dried powder 3 to 10 g of the mycelium culture medium, and then suspended well, followed by centrifugation at 5,000 rpm to 10,000 rpm for 10 to 30 minutes to obtain a supernatant. The extraction solvent corresponding to 2 to 3 times can be added and placed in a refrigerator at 1 to 5 ° C. for 10 to 15 hours. After only the supernatant is again centrifuged at 5,000 rpm to 10,000 rpm for 10 minutes to 30 minutes in the stationary material, the precipitate may be recovered to prepare a crude extracellular polysaccharide. The crude extracellular polysaccharide may be vacuum-dried at 30 ° C. or lower to obtain extracellular polysaccharide.

The extraction solvent may be a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, acetone, ether, chloroform and ethyl acetate or a mixed solvent thereof, and more specifically, water, methanol, ethanol, butanol It may be a solvent selected from the group consisting of acetone and ethyl acetate or a mixed solvent thereof, and more specifically, may be water or 50 to 80% (v / v) ethanol aqueous solution.

The pharmaceutical composition of the present invention exhibits the effect of preventing or eliminating hangovers by reducing blood ethanol and / or acetaldehyde concentration.

The extracellular polysaccharide may be included in 0.1 to 80% by weight, specifically, 0.1 to 50% by weight relative to the total weight of the pharmaceutical composition. In addition, the pharmaceutical composition may appropriately include a mycelium culture medium, a dry powder thereof, or an extract of the mycelium culture medium of Seriphoria laccerata in an amount corresponding to the content of the extracellular polysaccharide. However, the effective amount of the extracellular polysaccharide, the culture solution containing the same, the dry powder of the culture solution or the extract of the culture solution may be appropriately adjusted according to the method of use and purpose of the pharmaceutical composition.

The pharmaceutical composition is an extracellular polysaccharide produced by Seriphoria laccerata, mycelium culture medium of the Seriphoria laccerata, containing the dry powder of the mycelium culture medium or extract of the mycelium culture medium as an active ingredient In addition, it may further include suitable carriers, excipients and diluents commonly used in the pharmaceutical art.

The pharmaceutical compositions according to the invention may be formulated according to conventional methods. Suitable formulations include, but are not limited to, tablets, pills, powders, granules, dragees, hard or soft capsules, solutions, suspensions or emulsions, injections, suppositories, and the like.

The pharmaceutical compositions according to the invention can be prepared in suitable formulations using pharmaceutically inert organic or inorganic carriers. That is, when the formulation is a tablet, coated tablet, dragee and hard capsule, it may include lactose, sucrose, starch or derivatives thereof, talc, calcium carbonate, gelatin, stearic acid or salts thereof. In addition, when the formulation is a soft capsule, it may include polyols of vegetable oils, waxes, fats, semisolids and liquids. Furthermore, when the formulation is in the form of a solution or syrup, it may include water, polyols, glycerol, and / or vegetable oils and the like.

The pharmaceutical composition according to the present invention may further include a preservative, a stabilizer, a humectant, an emulsifier, a solubilizer, a sweetener, a colorant, an osmotic agent, an antioxidant, and the like in addition to the carrier.

The method of administering the pharmaceutical composition according to the present invention can be easily selected according to the dosage form, and can be administered orally or parenterally. The dosage may vary depending on the age, sex, weight, severity, and route of administration of the patient, but generally 5 to 1,000 mg per kilogram of body weight, specifically 10 to 600, based on the extracellular polysaccharide as an active ingredient. The mg may be administered once to three times daily. However, the above dosage does not limit the scope of the present invention.

The pharmaceutical composition according to the present invention not only provides excellent hangover relieving effect but also has little toxicity and side effects due to the drug, so that it can be used with confidence even during long-term use.

In addition, the present invention is an extracellular polysaccharide produced by Seriphoria laccerata, mycelium culture medium of the Seriporia laccerata containing the extracellular polysaccharide, the dry powder of the mycelium culture medium or the extract of the mycelium culture medium active ingredient It provides a health functional food for preventing or improving the hangover.

The extracellular polysaccharide, mycelium culture medium of the three liporia laccerata containing the same, its dry powder or extract is as described above.

The dietary supplement according to the present invention may be in the form of a powder, granule, tablet, capsule or beverage. In addition, the health functional food according to the present invention may be candy, chocolate, gum, tea, vitamin complex, health supplements and the like.

At this time, the content of the extracellular polysaccharide according to the invention contained in the health functional food, mycelium culture medium containing the same, the dry powder of the mycelium culture medium or the extract of the mycelium culture medium, 0.01 to 50% by weight relative to the total weight of the health food %, Specifically 0.1 to 20% by weight, and in the case of a beverage, it may be included in an amount of 0.02 to 10 g, specifically 0.3 to 1 g, based on 100 ml of the beverage.

The dietary supplement is an extracellular polysaccharide of the present invention, a food supplement that is acceptable in food supplements with a mycelium culture medium of Seriphoria laccerata containing the extracellular polysaccharide, a dry powder of the mycelium culture medium or an extract of the mycelium culture medium. It may further include.

The health functional food may have an excellent hangover relief effect. Health functional food according to the present invention can be ingested with confidence even in the long-term taking as well as the above-described effects, there is little toxicity and side effects.

In addition, the extracellular polysaccharide produced by the seriporia laccerata of the present invention, the mycelium culture medium of the seriporia laccerata containing the same, the dry powder of the mycelium culture medium or the extract of the mycelium culture solution is hangover prevention and / or hangover It can be used as an additive in various medicines, quasi-drugs and foods for the purpose of showing the relieving effect, wherein the amount and the use form can be appropriately adjusted according to the purpose.

In addition, the present invention is an extracellular polysaccharide produced by the above-mentioned Ceriporia laccerata, mycelium culture medium of the Ceriporia laccerata containing the extracellular polysaccharide, dry powder of the mycelium culture medium or the mycelium culture medium Provided are methods for preventing or eliminating hangovers comprising administering an extract to a subject in need thereof.

The subject may be a mammal, specifically a human.

Furthermore, the present invention is an extracellular polysaccharide produced by the above-mentioned Ceriporia laccerata, for use in the preparation of a hangover prevention or remedy composition, of the Ceriporia laccerata comprising the extracellular polysaccharide A mycelium culture solution, a dry powder of the mycelium culture solution or an extract of the mycelium culture solution is provided.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

EXAMPLE

Preparation Example 1. Mycelium culture solution of Ceriporia laccerata, dry powder thereof, extract and Preparation of extracellular polysaccharide (hereinafter referred to as "EPS")

1.1 Serifolia Lacserata  Preparation of Mycelia Culture

The mycelium of Seriphoria laccerata isolated from oak reimbursement in Sangju, Gyeongbuk was grown by subculture and stored frozen at -80 ℃. The cryopreserved strain was passaged 2-3 times in PDA (Potato dextrose agar) medium (87 plastic cultures; Difco, Becton Dickinson and Company) and stored the strain (hereinafter referred to as "PDA culture strain") in a 4 ° C refrigerator. Was used. Then, after preparing 600 ml of PDB (Potato dextrose broth) medium (Difco, Becton Dickinson and Company) in an Erlenmeyer flask, one PDA culture strain was added thereto, followed by shaking culture at 25 ° C. for 8 days to obtain a PDB culture strain.

On the other hand, 1.5% by weight of sugar, 0.5% by weight of glucose, 0.5% by weight of potato starch, 0.25% by weight of water, 0.25% by weight of soybean meal, 0.75% by weight of soy flour, 0.05% by weight of magnesium sulfate (MgSO ₄ ) Injecting the liquid culture medium containing 0.05% by weight of potassium monophosphate (KH ₂ PO ₄ ), 0.05% by weight of potassium diphosphate (K ₂ HPO ₄ ) and the remaining amount of water into an 800 L fermenter at 1.5 kgf / cm 2. Sterilization at 121 ° C. for 20 minutes. Thereafter, 600 ml of the PDB culture strain was inoculated with a starter while cooled to 23 ° C., and the air was aerated at 0.5 to 1.5 kgf / cm 2. The light source was maintained at a blue LED, the illuminance was 0.5 LUX, and the concentration of carbon dioxide was 2,000 ppm. The mycelium culture medium of Ceriporia laccerata was prepared by liquid culture of the Ceriporia laccerata mycelium at a constant temperature of 23 ° C. for 10 days.

1.2 Serifolia Lacserata  Preparation of dry powder of mycelium culture

The mycelium culture medium of Ceriporia laccerata prepared in Preparation Example 1.1 was vacuum-dried and powdered by vacuum freeze drying at 25 ° C. for 72 hours using a vacuum freeze dryer to prepare a dry powder of the mycelium culture medium of Ceriporia laccerata.

1.3 Serifonia Lacserata  Preparation of extract of mycelium culture

100 ml of distilled water was added to 5 g of the dry powder of the mycelium culture medium of Ceriporia laccerata prepared in Preparation Example 1.2, and then suspended well, and centrifuged at 8,000 rpm for 20 minutes. Ethanol corresponding to 2-3 times the volume of the supernatant was added to its supernatant and left to stand at 4 ° C. for 12 hours. Then, the supernatant was taken from the stationary material to prepare an extract of the mycelium culture medium of Seriporia laccerata.

1.4 Serifonia Lacserata  Preparation of EPS from Extracts of Mycelium Cultures

The extract of the mycelium culture medium of Seriphoria laccerata prepared in Preparation Example 1.3 was further centrifuged at 8,000 rpm for 20 minutes, and the precipitate was recovered to obtain a crude EPS. The crude EPS was vacuum-dried at 25 ° C. for 72 hours using a vacuum freeze dryer to obtain EPS produced by Ceriporia laccerata.

Example 1 Evaluation of Characteristics of EPS

1.1 Gel Permeation Chromatography, GPC Molecular weight of EPS using

The EPS prepared in Preparation Example 1 was dissolved in 0.1 M Na ₂ SO ₄ /0.05 M NaN ₃ (adjust the pH to 4 with glacial acetic acid) to 1% (w / v) and then at 4,000 rpm. After centrifugation for 0.5 hour, only the supernatant was filtered with a 0.45 μm syringe filter and analyzed by GPC.

Specifically, the GPC analysis conditions using the refractive index as a detector, GPC column using a OHpak SB 805 HQ (Shodex, Japan) 0.1 M Na ₂ SO ₄ /0.05 M NaN ₃ (mobile pH to pH 4 as glacial acetic acid) ), And the flow rate of the mobile phase was flowed at a rate of 1.0 mL / minute. Standard curves were prepared using Dextran (American Polymer Corporation, USA) with different molecular weights (130 kDa, 400 kDa, 770 kDa, or 1200 kDa), and Knauer K-2310 (refractive index (RI) measuring instrument). Germany) to determine the molecular weight of the EPS. The measurement conditions are summarized in Table 1 below.

	Measurement of molecular weight
GPC system	Knauer K-501 System
column	OHpak SB 805 HQ (Shodex, Japan)
Mobile phase	0.1 M Na 2 SO 4 /0.05 M NaN 3 / pH 4
Flow rate	1.0 ml / min
Measuring instrument	RI (Knauer K-2310)

As a result, the molecular weight of EPS of the present invention was found to be about 120 kDa.

1.2 Determination of sugar and protein content in EPS

The EPS prepared in Preparation Example 1 was second-purified and treated with proteolytic enzymes to determine sugar and protein content.

Specifically, the super purified EPS (EPS prepared in Preparation Example 1) was dissolved in distilled water and centrifuged at 8,000 rpm for 20 minutes to separate the supernatant. Ethanol corresponding to 2-3 times the volume of the supernatant was added to the separated supernatant and placed in a 4 ° C. refrigerator for 12 hours. Thereafter, only the supernatant was again centrifuged at 8,000 rpm for 20 minutes in the stationary material, and the precipitate was recovered to obtain a second purified EPS. The second purified EPS was dissolved in distilled water, and the proteolytic enzyme alcalase was treated at 50 ° C. for 30 minutes at a concentration of 0.5% (w / v).

Sugar content was measured using the phenol-sulfuric acid method. Specifically, 25 μl of 80% (w / v) phenol was added to 1 mL of the sample diluted by concentration, and then 2.5 mL of sulfuric acid was added and cooled to room temperature. The absorbance was measured at 465 nm to calculate the sugar content.

In addition, the protein content was measured by the BCA method (Smith PK et al. , Analytical Biochemistry , 150 (1): 76-85, 1985), and bovine serum albumin was used as a standard.

The sugar content and protein content measured as described above are shown in Table 2 below, and the sugar content was found to be 45 to 51 wt% and the protein content was 33 to 34 wt%.

	yield(%)	Total sugar content (%)	Total protein content (%)
EPS	1.22 ± 0.03	45.32 ± 1.41	34.17 ± 0.73
Secondary Refined EPS	0.78 ± 0.01	50.49 ± 0.52	33.50 ± 2.79
Enzyme-treated EPS *	0.24 ± 0.06	51.39 ± 1.32	34.61 ± 1.51

^* Enzyme treatment: 0.5% Al Calle Izu, 50 ℃, 30 minutes.

Each figure is mean ± SE (n≥3).

In addition, the analysis of sugar components of EPS revealed that EPS mainly contains mannose, galactose and glucose.

Example 2 Verification of Hangover Elimination Effect in Mouse

In order to evaluate the hangover resolution of the Ceriporia laccerata mycelium culture medium of Preparation Example 1.1, the mice were treated with ethanol, treated with various concentrations of the Ceriphoria laccerata mycelium culture medium, and the concentration of blood alcohol in the mouse was measured. It was.

Specifically, a 7-week-old C57BL / 6J mouse lit and turned off at 12-hour intervals under conditions of temperature 21 ± 3 ° C, relative humidity 50 ± 5%, and illuminance 200 to 300 lux while freely ingesting solid feed and tap water. After adaptation, only healthy animals were selected and used for testing. After weighing the subjects who had no health problems during the adaptation period, 50 animals near the mean weight were selected. The selected animals were divided into 5 groups of 10 animals by random selection method, fasted 18 hours before the experiment, and water was supplied only. Thereafter, blood alcohol concentration was measured according to the administration time.

Among the fasted mice, the first group received only distilled water as a normal control group, and the second group received oral administration of 40% (v / v) ethanol at an amount of 5 g / kg body weight as a negative control group, and the third group as an experimental group. 40% (v / v) ethanol and 100 mg / kg body weight of 5 g / kg body weight were orally administered with the culture medium of Ceriporia laccerata of Preparation Example 1.1, and the fourth group was 5 g as the experimental group. 40% (v / v) ethanol and 500 mg / kg body weight of the amount of the weight of the weight of the culture medium of Ceriporia lacserata of Preparation Example 1.1 was administered orally. Before oral administration, immediately after oral administration, 1.5 ml of blood was collected by cardiac sampling of control and experimental mice after 30, 120 and 300 minutes of oral administration. Blood ethanol concentration (mM) was analyzed from the collected blood using Headspace-GC / MS (Perkin Elmer, clarus 600T), and the results are shown in FIG. 1.

As shown in Figure 1, 30 minutes after the ethanol administration, the blood ethanol concentration was 10.76 and 3, respectively, the group 3 and the fourth group added to the culture medium of the seriporia lacserata mycelium compared to the second group administered only ethanol, respectively. % And 7.03% lower. In addition, after 300 minutes of ethanol administration, the third and fourth groups were 54.46% and 84.04% lower than the second group to which only ethanol was administered.

Further, as a result of calculating the area under the curve (AUC) in FIG. 1, the AUC of the second group was 1398.9 mM × min, the AUC of the third group was 1238.7 mM × min, and the AUC of the fourth group was 1195.05. mM × min. As a result, it was found that blood ethanol concentrations of the third and fourth groups were 11.45% and 14.57%, respectively, compared to the second group administered only ethanol.

This suggests that the mycelium culture medium of the seriporia laccerata of the present invention reduces the concentration of ethanol in the blood, and thus the mycelial culture solution of the seriporia laccerata has a hangover effect.

Example 3. Clinical Trials

In order to evaluate the hangover resolution effect of the dry powder of mycelium culture medium of Ceriporia laccerata of Preparation Example 1.2, blood ethanol concentration (%) was measured and clinical questionnaire was performed after drinking. Age, gender composition for the panel of the experiment is shown in Table 3 below.

Panel composition	Medicine		Placebo
	male	female	male	female
29 years old or younger	0	0	0	One
30 to 34 years	2	0	2	0
35 to 39 years old	0	0	0	0
40 years old or older	5	0	3	0
system	7	0	5	One

Specifically, for the panel ingesting ethanol in an amount of 1.5 g / kg body weight, the drug group is 90.91% by weight dry powder of Ceriporia laccerata mycelium culture medium of Preparation Example 1.2, 6.49% by weight crystalline cellulose, silicon dioxide 1.8 Two tablets (550 mg / tablet) made of a mixture of wt% and 0.8 wt% magnesium stearate were administered orally, and the placebo group was a tablet made of a mixture of lactose (550 mg / tablet) instead of dry powder of Ceriporia laccerata mycelia. ) Was administered orally. The blood alcohol concentration of the panel was measured immediately after drinking, 30 minutes after, 60 minutes after, 120 minutes and after 16 hours, and the measurement was performed using a breathalyzer measuring blood levels by expiration. In addition, 16 hours after drinking, the panel was subjected to a clinical questionnaire. The relative concentration (%) of blood ethanol measured as a reference (100%) immediately after drinking is shown in Table 4 and FIG. 2, and the average value of the clinical questionnaire results after drinking 16 hours is shown in Table 5 below.

	Immediately after drinking	30 minutes after drinking	60 minutes after drinking	After 120 minutes	16 hours after drinking
Average Blood Ethanol Relative Concentration in the Drug Group (7) (%)	100	92.31	76.92	60	0
Relative Concentrations of Average Blood Ethanol in the Placebo Group (6)	100	91.53	100	66.1	0

As shown in Table 4 and FIG. 2, the blood ethanol concentration after drinking was 23.08% and 9.23% after 60 minutes and 120 minutes of drinking in the medicament group administered the culture medium of the seriporia laccerata mycelium of the present invention compared to the placebo group. Low.

Further, as a result of calculating the area under the curve (AUC) of FIG. 2, the AUC of the drug group was 584.62 relative% x time, and the AUC of the placebo group was 645.75 relative% x time. Through this, it was confirmed that the blood alcohol concentration of the drug group compared to the placebo group was reduced by 9.47%.

This shows that the seriporia laccerata mycelium culture of the present invention has a hangover-relieving activity by reducing the absorption of ethanol in the blood.

	Hangover-related symptoms					Fatigue-related symptoms			Arousal-related symptoms		Digestive symptoms		Headache-related symptoms
	Memory break	Rumble	tinnitus	Vomiting / Nausea	thirst	Sweat	Helplessness	drowsiness	Weather difficulties	Poor concentration	diarrhea	colic	headache	whirl
Jinyaeng-gun	One	1.14	One	One	1.43	One	1.14	1.14	1.14	1.14	1.29	One	1.14	1.29
Placebo group	One	1.5	One	One	2.33	One	1.67	2	1.83	1.67	1.17	1.5	1.83	1.67
Symptom improvement compared to placebo	-	24%	-	-	38.63%	-	31.74%	43%	37.7%	31.74%	-	33.33%	37.7%	22.75%

In Table 5, 1 is no or little obstacle in life, 2 is little or very weak in life, 3 is no problem in life but is slightly severe, and 4 is slightly disturbed or generally severe in life. For example, 5 indicates that the symptoms are severely disturbed or very severe in life.

As shown in Table 5, compared with the placebo group, the Jinja group had direct hangover-related symptoms (24% relief and 39% relief from thirst), fatigue-related symptoms (32% relief of helplessness and 43% relief of drowsiness) due to hangover, and hangover. Arousal-related symptoms (relieves 38% of difficulty in weathering and 32% reduction in concentration), digestive-related symptoms from hangovers (33% relief from abdominal pain) and headache-related symptoms from hangovers (relieves headaches 38% and dizziness 23%) Improved.

Through the above results, it was found that the culture medium of the seriporia laccerata mycelium of the present invention has a hangover relief effect.

Claims (21)

1. Extracellular polysaccharide produced by Ceriporia lacerata ( Ceriporia lacerata ), mycelium culture medium of the seriporia laccerata containing the extracellular polysaccharide, dry powder of the mycelium culture solution or extract of the mycelium culture medium as an active ingredient A pharmaceutical composition for preventing or eliminating hangovers.
2. The method of claim 1,

   The extracellular polysaccharide comprises a 40 to 60% by weight of sugar and 30 to 40% by weight of protein and has a molecular weight of 100 to 150 kDa, hangover prevention or remedy pharmaceutical composition.
3. The method of claim 2,

   The extracellular polysaccharide comprises 43 to 47% by weight of sugar and 33 to 36% by weight of protein and has a molecular weight of 115 to 125 kDa, hangover prevention or remedy pharmaceutical composition.
4. The method of claim 2,

   The sugar comprises mannose, galactose and glucose, pharmaceutical composition for preventing or alleviating hangovers.
5. The method of claim 1,

   The extracellular polysaccharide is

   (a) liquid culture of the mycelium of the seriporia laccerata to prepare a mycelium culture of the seriporia laccerata;

   (b) drying and powdering the mycelium culture medium of Ceriporia laccerata; And

   (c) extracting the dry powder of the mycelium culture medium of the seriporia laccerata with a solvent, and then filtering and concentrating under reduced pressure, a pharmaceutical composition for preventing or eliminating hangovers.
6. The method of claim 5,

   The medium for culturing the liquid is sugar, glucose, starch, hydrated water, soybean meal, soybean meal, magnesium sulfate (MgSO ₄ ), potassium monophosphate (KH ₂ PO ₄ ), potassium diphosphate (K ₂ HPO ₄ ) and water To include, the hydrogen ion concentration is pH 4.5 ~ 6.0, hangover prevention or remedy pharmaceutical composition.
7. The method of claim 5,

   The liquid culture is carried out under a blue LED light source, and is carried out by maintaining the concentration of carbon dioxide at 1,000 to 2,000 ppm, hangover prevention or remedy pharmaceutical composition.
8. The method of claim 1,

   The extracellular polysaccharide is contained in an amount of 0.1 to 80% by weight based on the total weight of the composition, hangover prevention or remedy pharmaceutical composition.
9. The method of claim 1,

   The pharmaceutical composition for reducing or reducing the concentration of ethanol or acetaldehyde in the blood, hangover prevention or pharmaceutical composition.
10. Hangover containing an extracellular polysaccharide produced by Seriphoria laccerata, mycelium culture medium of the Seriphoria laccerata containing the extracellular polysaccharide, a dry powder of the mycelium culture medium or an extract of the mycelium culture medium as an active ingredient. Health functional food for prevention or improvement.
11. The method of claim 10,

    The extracellular polysaccharide comprises 40 to 60% by weight of sugar and 30 to 40% by weight of protein and has a molecular weight of 100 to 150 kDa, hangover prevention or improvement health functional food.
12. The method of claim 11,

    The extracellular polysaccharide comprises 43 to 47% by weight of sugar and 33 to 36% by weight of protein and has a molecular weight of 115 to 125 kDa, hangover prevention or improvement health functional food.
13. The method of claim 11,

    The sugar containing mannose, galactose and glucose, health functional food for preventing or improving hangover.
14. The method of claim 10,

    The extracellular polysaccharide is

    (a) liquid culture of the mycelium of the seriporia laccerata to prepare a mycelium culture of the seriporia laccerata;

    (b) drying and powdering the mycelium culture medium of Ceriporia laccerata; And

    (c) extracting the dry powder of the mycelium culture medium of Seriphoria laccerata with a solvent and then filtering and concentrating under reduced pressure, a health functional food for preventing or improving hangover.
15. The method of claim 14,

    The medium for culturing the liquid is sugar, glucose, starch, hydrated water, soybean meal, soybean meal, magnesium sulfate (MgSO ₄ ), potassium monophosphate (KH ₂ PO ₄ ), potassium diphosphate (K ₂ HPO ₄ ) and water To include, the hydrogen ion concentration is pH 4.5 ~ 6.0, health functional food for preventing or improving hangover.
16. The method of claim 14,

    The liquid culture is carried out by maintaining the concentration of carbon dioxide under a blue LED light source at 1,000 ~ 2,000 ppm, hangover prevention or improvement health functional food.
17. The method of claim 10,

    The health functional food is in the form of a powder, granules, tablets, capsules or drinks, health functional food for preventing or improving a hangover.
18. The method of claim 10,

    The health functional food is candy, chocolate, gum, tea, vitamin complex or health supplement food, health functional food for preventing or improving hangover.
19. The extracellular polysaccharide produced by Seriphoria laccerata, the mycelium culture medium of the Seriphoria laccerata containing the extracellular polysaccharide, the dry powder of the mycelium culture medium or the extract of the mycelium culture solution are administered to a subject in need thereof. Hangover prevention or cancellation method including thing to do.
20. The method of claim 19,

    A method for preventing or alleviating a hangover, wherein the subject is a mammal.
21. Extracellular polysaccharide produced by Seriphoria laccerata, Mycelium culture medium of Seriphoria laccerata containing the extracellular polysaccharide, Dry powder of the mycelium culture medium for use in the preparation of hangover prevention or remedy composition Or the use of an extract of the mycelium culture.

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